title: cAMP inhibits migration, ruffling and paxillin accumulation in focal adhesions of pancreatic ductal adenocarcinoma cells: effects of PKA and EPAC.
abstract: We demonstrated that increasing intracellular cAMP concentrations result in the inhibition of migration of PANC-1 and other pancreatic ductal adenocarcinoma (PDAC) cell types. The rise of cAMP was accompanied by rapid and reversible cessation of ruffling, by inhibition of focal adhesion turnover and by prominent loss of paxillin from focal adhesions. All these phenomena develop rapidly suggesting that cAMP effectors have a direct influence on the cellular migratory apparatus. The role of two primary cAMP effectors, exchange protein activated by cAMP (EPAC) and protein kinase A (PKA), in cAMP-mediated inhibition of PDAC cell migration and migration-associated processes was investigated. Experiments with selective activators of EPAC and PKA demonstrated that the inhibitory effect of cAMP on migration, ruffling, focal adhesion dynamics and paxillin localisation is mediated by PKA, whilst EPAC potentiates migration.
authors.affiliation: Department of Cellular and Molecular Physiology, The University of Liverpool, Crown Street, Liverpool L69 3BX, UK.
authors.forename: Alex
authors.initials: A
authors.lastname: Burdyga
authors.affiliation: Department of Cellular and Molecular Physiology, The University of Liverpool, Crown Street, Liverpool L69 3BX, UK.
authors.forename: Alan
authors.initials: A
authors.lastname: Conant
authors.affiliation: Department of Cellular and Molecular Physiology, The University of Liverpool, Crown Street, Liverpool L69 3BX, UK.
authors.forename: Lee
authors.initials: L
authors.lastname: Haynes
authors.affiliation: The Johns Hopkins University School of Medicine, Department of Neuroscience, 725 North Wolfe Street, Baltimore, MD 21205, USA.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: The Netherlands Cancer Institute, Cell Biology I, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
authors.forename: Kees
authors.initials: K
authors.lastname: Jalink
authors.affiliation: NIHR Liverpool Pancreas Biomedical Research Unit, The University of Liverpool, Crown Street, Liverpool L69 3BX, UK.
authors.forename: Robert
authors.initials: R
authors.lastname: Sutton
authors.affiliation: Department of Molecular and Clinical Cancer Medicine, 6th Floor, Duncan Building, Daulby Street, Liverpool, L69 3GA, UK.
authors.forename: John
authors.initials: J
authors.lastname: Neoptolemos
authors.affiliation: NIHR Liverpool Pancreas Biomedical Research Unit, The University of Liverpool, Crown Street, Liverpool L69 3BX, UK; Department of Molecular and Clinical Cancer Medicine, 6th Floor, Duncan Building, Daulby Street, Liverpool, L69 3GA, UK.
authors.forename: Eithne
authors.initials: E
authors.lastname: Costello
authors.affiliation: Department of Cellular and Molecular Physiology, The University of Liverpool, Crown Street, Liverpool L69 3BX, UK. Electronic address: a.tepikin@liv.ac.uk.
authors.forename: Alexei
authors.initials: A
authors.lastname: Tepikin
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000230:Adenocarcinoma / Q000201:enzymology / Q000473:pathology; D021441:Carcinoma, Pancreatic Ductal / Q000201:enzymology / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D022081:Cell Surface Extensions / Q000187:drug effects / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D006801:Humans; D009361:Neoplasm Invasiveness; D010190:Pancreatic Neoplasms / Q000201:enzymology / Q000473:pathology; D051419:Paxillin / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects
chemical_list: D020662:Guanine Nucleotide Exchange Factors; D051419:Paxillin; C116122:RAPGEF3 protein, human; D005576:Colforsin; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D015056:1-Methyl-3-isobutylxanthine
keywords: 3-isobutyl-1-methylxanthine; 6Bnz-cAMP; 8, 8-(4-chlorophenylthio)-2′-O-methyl-cAMP; 8, 8-bromoadenosine-cAMP; 8Br-cAMP; 8pCPT; A-kinase activity reporter four; AKAR4; CFP(nd)-EPAC(dDEP/CD)-Venus(d); Cell migration; DMEM; Dulbecco's modified Eagle's medium; EPAC; FBS; Frsk; H84; IBMX; N6-benzoyl-cAMP; PBS; PDAC; PKA; Pancreatic ductal adenocarcinoma; Paxillin; cAMP; exchange protein activated by cAMP; foetal bovine serum; forskolin; pancreatic ductal adenocarcinoma; phosphate buffered solution; protein kinase A